Composition for preventing or treating dentin-associated symptoms or diseases, and method using the same

ABSTRACT

Provided is a formulation for oral teeth, which includes a plurality of calcium ion carriers and a plurality of calcium-containing particulates. The particulates are carried by the calcium ion carriers, such that the formulation can prevent or rapidly treat dentin-associated symptoms or diseases, while providing a prolonged prophylactic or therapeutic effect.

BACKGROUND OF THE INVENTION 1. Field of the Invention

The present invention relates to a formulation for use in oral teeth,and more particularly relates to a formulation for propylaxising andtreating dentin-associated diseases or symptoms.

2. Description of Associated Art

Dentin, which is also known as “dentine,” is a tissue constituting thetooth matrix, and located between dental enamel and dental pulp. Dentinis composed of 70% of inorganic substance, 20% of organic substance and10% of water. The hardness of dentin is lower than that of dentalenamel, but higher than that of cementum. Dentinal tubules throughoutthe dentin are arranged radially from the surface of the dental pulptowards the dental enamel. The dentinal tubules are wider at the endsnear the dental pulp. The closer toward the surface the dentinal tubuleswith branches therebetween are, the narrower they are.

Common dentin-associated diseases or symptoms which cause pain, includesdental caries, tooth wearing, enamel loss and dentin hypersensitivityetc.

Dentin hypersensitivity is sometimes referred to as “toothhypersensitivity” and “hypersensitive dentine.” Currently, various typesof products or methods for relieving dentin hypersensitivity have beendeveloped. However, up to now, none of the products or methods canprovide rapid and long-term relieving effect.

Generally, the clinical methods for treating dentin hypersensitivity canfall into the following two categories: (1) chemical desensitizingmethod; and (2) physical desensitizing method.

As to the chemical desensitizing methods, corticosteroids were used inthe early years to suppress inflammation. However, such methods areineffective. Further, protein precipitation, which is also categorizedas a chemical desensitizing method, utilizes a chemical agent tocoagulate and denature the proteins in the dentinal tubule. For example,a formulation containing silver nitrate, phenol, formaldehyde orstrontium chloride is used to denature collagen, and then formsprecipitates which block the openings of the dentinal tubules. However,such formulation stimulates dental pulp and gingival, and the relapserate is extremely high. Further, silver nitrate dyes teeth blackpermanently.

Moreover, chemical desensitizing methods also include a treating methodfor paralyzing pulp nerves. For example, some commercially availabledesensitizing toothpastes use potassium nitrate to suppress theexcitation of pulp nerves. However, the clinical cases have shown thatthe pain on a patient cannot be relieved until the desensitizingtoothpaste is persistently used for two weeks, and the therapeuticeffect can last for only several months. That is to say, the methods forparalyzing pulp nerves cannot provide rapid and long-term therapeuticeffects. Also, the long-term use of potassium nitrate leads to disordersassociated to the paralysis of pulp nerves.

On the other hand, as to the physical desensitizing methods, forexample, a sealant for dentinal tubules is used to directly seal theopenings of dentinal tubules. The sealant includes, for example, resins,glass ionomer cements and or the like. For example, Jensen et al. (“Acomparative study of two clinical techniques for treatment of rootsurface hypersensitivity,” Gen. Dent. 35:128-132.) proposed a method fordirectly sealing the openings of dentinal tubules using a resin-typedentin bonding agent. Although this method can immediately relive thepain caused by dentin hypersensitivity, it cannot provide long-termtherapeutic effects. More specifically, the clinical cases have shownthat, after a 6-months treatment, the resin-type bonding agent detachessignificantly from the surfaces of teeth. As to the glass ionomercements, Low et al. (“The treatment of hypersensitive cervical abrasioncavities using ASAP cement,” J. Oral Rehabil. 8(1):81-9) used glassionomer cements to treat dentin hypersensitivity in 1981. Although glassionomer cements can provide therapeutic effects, this type of materialwill be removed by constantly brushing the tooth. Further, Hansen et al.(“Dentin hypersensitivity treated with a fluoride-containing varnish ora light-curd glass ionomer liner,” Scand. J. Dent. Res. 100(6):305-9)used resin-enhanced glass ionomer cements to treat dentinhypersensitivity, but still no long-term therapeutic effects wereachieved.

Accordingly, it is an urgent and important issue to provide rapid andprolonged effects to relieve dentin-associated symptoms and diseases.

SUMMARY OF THE INVENTION

In view of the foregoing, the present invention provides a formulationfor oral teeth, comprising a plurality of calcium ion carriers and aplurality of calcium-containing particulates, wherein thecalcium-containing particulates are carried by the calcium ion carriers.Therefore, the formulation for oral teeth of the present invention canprevent or rapidly treat dentin-associated symptoms or diseases, and canprovide prolonged prophylactic or therapeutic effects.

The present invention further provides a method for preventing ortreating a dentin-associated symptom or disease, comprising a step ofadministering the aforesaid formulation for oral teeth of the presentinvention to the oral cavity of a subject.

The present invention further provides a method for dental therapy,comprising the steps of: providing a composition comprisingcalcium-containing particulates, calcium- and phosphorus-containingparticulates, fluoro-containing particulates or a combination thereofand microparticles; mixing the composition and an acidic solution toform a formulation; and administering the formulation to the oral cavityof a subject.

The present invention further provides a composition for oral care,comprising a plurality of microparticles, wherein each of themicroparticles has a plurality of pores, and the pores have a porediameter ranging from 1 to 100 nm; and a calcium-containing component, aphosphorus-containing component, a calcium- and phosphorus-containingcomponent, a fluoro-containing or a combination thereof. Thereby, thecomposition for oral care of the present invention is beneficial to theprevention or rapid treatment of a dentin-associated symptom or disease,and can provide prolonged prophylactic or therapeutic effects.

Moreover, the present invention provides a method for preventing ortreating a dentin-associated symptom or disease, comprising the step ofadministering the aforesaid formulation for oral care the presentinvention to the oral cavity of a subject.

In addition, the present invention provides a product comprising theaforesaid formulation for oral care of the present invention. Theproduct comprises dental products, toothpastes, tooth powder, ointments,chewing gums, troches, mouthwash, toothbrushes or strips.

BRIEF DESCRIPTION OF THE DRAWINGS

The present invention can be more fully understood by reading thefollowing detailed description of the preferred embodiments, withreference made to the accompanying drawings, wherein:

FIG. 1 shows an SEM observation of the Ca—Si-Based porous particles ofan embodiment of the present invention;

FIG. 2a shows an SEM observation of a dental test slice coated with theCaCO₃@porous SiO₂ formulation of an embodiment of the present invention;

FIG. 2b shows an SEM observation of a dental test slice coated with theSeal & Protect® of a comparative example of the present invention;

FIG. 3a shows an SEM observation of a dental test slice of a live dogcoated with the CaCO₃@porous SiO₂ formulation of an embodiment of thepresent invention;

FIG. 3b shows an SEM observation of a dental test slice of a live dogcoated with the Seal & Protect® of a comparative example of the presentinvention;

FIG. 4a shows an observation of the pulp tissue morphology of a dentaltest slice of a live dog coated with the CaCO₃@porous SiO₂ formulationof an embodiment of the present invention; and

FIG. 4b shows an observation of the pulp tissue morphology of a dentaltest slice of a live dog coated with the Seal & Protect® of acomparative example of the present invention.

DETILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The following specific examples are used to exemplify the presentinvention. A person of ordinary skills in the art can conceive the otheradvantages of the present invention, based on the disclosure of thespecification of the present invention. The present invention can alsobe implemented or applied as described in different specific examples.It is possible to modify and or alter the above examples for carryingout this invention without contravening its spirit and scope, fordifferent aspects and applications.

In order to solve the issues in prior art, the inventors the presentinvention accomplished a formulation for oral teeth after performing avariety of experiments. The formulation comprises a calcium ion carrierand a calcium-containing particulate, wherein the calcium-containingparticulate is carried by the calcium ion carrier.

The form the formulation for oral teeth of the present inventionpresents is not limited. Preferably, the formulation for oral teeth ofthe present invention exists as a gel, a paste, a slurry, an emulsion ora glue.

The carrier of the calcium ion of the present invention is not limited,but it may be composed of an organic substance or an inorganicsubstance. More specifically, in some preferred embodiments, the organicsubstance constituting the calcium ion carrier is preferably a polymer,and more preferably a porous polymer. In some preferred embodiments, theorganic substance constituting the calcium ion carrier may beester-based polymer or alkene-based polymer. For example, the preferredester-based polymers may include acrylic latex or polystyrene, and thepreferred alkene-based polymer may include nylon. Further, the calciumion carrier may be composed of a combination of acrylic latex,polystyrene and nylon.

As to the inorganic substance constituting a calcium ion carrier, insome preferred embodiments, the organic substance includes an oxide. Theoxide constituting the calcium ion carrier is not limited, butpreferably silicon dioxide, silicoaluminate, titanium dioxide, zincoxide, aluminum oxide or a combination thereof.

In a preferred embodiment of the formulation for oral teeth of thepresent invention, the calcium ion carrier is a microparticle having aplurality of pores. More specifically, the way the calcium ion carrierscarries the calcium-containing particulate is not limited. Preferably,the calcium-containing particulate is adhered to the pores of themicroparticles.

The calcium-containing particulate of the present invention is notlimited, but preferably composed of calcium oxide, calcium chloride,calcium carbonate, calcium nitrate, calcium hydroxide, calcium salt or acombination thereof. In some preferred embodiments, thecalcium-containing particulate may further comprise phosphorus. Morespecifically, the calcium-containing particulate comprising phosphorusis not limited, but preferably composed of calcium phosphate,hydroxyapatite, tricalcium phosphate, tetracalcium phosphate, CaHPO₄,Ca₈H₂(PO₄)₆.5H₂O, Ca₂P₂O₇, Na₂O—CaO—SiO₂—P₂O₅ or a combination thereof.

In a preferred embodiment of the formulation for oral teeth of thepresent invention, the pH value of the formulation is in a range of 2 to10, and preferably in a range of 7 to 10.

The aforesaid components of the formulation of the present invention areonly used to exemplary illustrate the proffered embodiment of thepresent invention, and not intended to limit the scope of the presentinvention. That is, the formulation for oral teeth of the presentinvention may further comprise a calcium-ion release agent, whichreleases calcium ions from the calcium-containing particulates carriedon the calcium-ion carries. More specifically, the calcium-ion releaseagent may be an acidic solution, and preferably at least one selectedfrom the group consisting of phosphoric acid, oxalic acid and citricacid, and more preferably phosphoric acid. The acidic solution may alsobe an organic acid or inorganic acid related or similar to any one ofphosphoric cid, oxalic acid and citric acid.

In a preferred embodiment of the formulation for oral teeth of thepresent invention, the concentration of the phosphoric acid is in arange of 1% to 65%, preferably in a range of 25% to 45%, and morepreferably in a range of 31% to 40%.

In another preferred embodiment of the formulation for oral teeth of thepresent invention, when the calcium-ion release agent is an acidicsolution, the ratio of the contents of the acidic solution and thecalcium ion carriers carrying calcium-containing particulates is in arange of 1 to 10 mL/g, preferably in a range of 2 to 5 mL/g.

As described above, the formulation for oral teeth of the presentinvention may optionally comprise or not comprise an acidic solution,which has a pH value in a range of 2 to 10. In an embodiment, the pHvalue may be in a range of 7 to 10. Alternatively, in the case ofcomprising the acidic solution, the pH value of the formulation may bein a range of 4 to 7, preferably in a range of 5 to 6.

Moreover, the formulation for oral teeth of the present invention mayfurther comprise components other than those mentioned above. Morespecifically, the formulation may further comprise a fluoro-containingcomponent, which may be a fluoro-containing compound, such as a fluoridesalt. Preferably, the fluoro-containing component is carried by theaforesaid calcium ion carrier. In some preferred embodiments, thefluoro-containing component is one selected from the group consisting ofammonium fluoride, calcium fluoride, sodium fluoride, potassiumfluoride, stannous fluoride, aluminum fluoride, sodiummonofluorophosphate, sodium hexafluorosilicate and a combinationthereof. Further, the formulation for oral teeth of the presentinvention may further comprise a pH stabilizer.

In another preferred example of the formulation for oral teeth of thepresent invention, the particulate diameters of the microparticles arein a range of 0.1 μm to 100 μm, and the pore diameters of the pores arein a range of 1 nm to 100 nm. Preferably, the pore diameters of thepores are in a range of 2 nm to 50 nm.

According to another aspect of the present invention, the presentinvention further provides a method for preventing or treating adentin-associated symptom or disease, comprising the step ofadministering the aforesaid formulation of the present invention to theoral cavity of a subject. More specifically, the dentin-associatedsymptoms which can be prevented or treated by the method of the presentinvention are not limited, and may include dentin hypersensitivity,crack tooth syndrome, enamel loss, dentin loss, cemento loss andpostoperative hypersensitivity. Enamel loss, dentin loss or cemento lossis generally caused by corrosion, abrasion, wearing or cracking to theteeth. Postoperative hypersensitivity generally occurs after dentaloperations such as teeth bleaching, prosthodontic or restoration.

On the other hand, the disease which can be prevented or treated by themethod of the present invention are not limited, preferably dentalcaries, root caries, tooth fracture, root fracture, cervical abrasion,tooth wearing or dentin-associated pulp disease.

According to another aspect of the present invention, the presentinvention provides a method for dental therapy, comprising the steps of:providing a composition comprising calcium-containing particulates,calcium- and phosphorus-containing particulates, fluoro-containingparticulates or a combination thereof and microparticles; mixing thecomposition and an acidic solution to form a formulation; andadministering the formulation to the oral cavity of a subject.

In a preferred embodiment of the method for dental therapy of thepresent invention, the microparticles each has a plurality of poreshaving pore diameters ranging from 1 to 100 nm. Calcium-containingparticulates, calcium- and phosphorus-containing particulates,fluoro-containing particulates or a combination thereof are adhered intothe plurality of the pores of the microparticles.

In another preferred embodiment of the method for dental therapy of thepresent invention, the aforesaid acidic solution is at least oneselected from the group consisting of a phosphoric acid, oxalic acid anda combination thereof. In this case, the concentration of the phosphoricacid may be in a range of 1% to 65%, preferably in a range of 25 to 45%,and more preferably in a range of 31% to 40%.

In another preferred embodiment of the method, in the step of formingthe formulation, the ratio of the contents of the acidic solution andparticles adhered with the calcium-containing particulates, the calcium-and phosphorus-containing particulates, the fluoro-containingparticulates or a combination thereof is in a range of 1 mL/g to 10mL/g. Preferably, in the step of forming the formulation, the ratio ofthe contents of the acidic solution and the calcium-containingparticulates, the calcium- and phosphorus-containing particulates, thefluoro-containing particulates or a combination thereof is in a range of2 mL/g and 5 mL/g. Further, in the step for forming the formulation, theformed formulation is preferably present as a gel, a paste, a slurry, anemulsion or a glue.

In another preferred embodiment, the method for dental therapy thepresent invention further comprises a step of administering an aqueoussolution to the oral cavity of the subject, and the pH value of theaqueous solution is greater than or equal to 7. The aqueous solutionadministered in the method for dental therapy the present invention isnot limited, preferably may be basic water or basic solution prepared inthe labor. With regard to the aforesaid basic water, it may be a basicwater obtained any source, such as a commercially available basic water,or basic water prepared in the labor. In addition, the aforesaid basicwater has a pH preferably greater than 7.

In another preferred embodiment, the method for dental therapy thepresent invention further comprises the step of administering water tothe oral cavity of the subject. Preferably, prior to the administeringof the alkaline solution, water is administered to the cavity of thesubject. More specifically, the number of times that water isadministered to the oral cavity of a subject is not limited, preferablyat least twice.

According to another aspect of the present invention, the presentinvention provides a composition for oral care, comprising a pluralityof microparticles, wherein each of the microparticles has a plurality ofpores, and the pores have a pore diameter ranging from 1 to 100 nm; anda calcium-containing component, a phosphorus-containing component, acalcium- and phosphorus-containing component, a fluoro-containingcomponent and a combination thereof.

In a preferred embodiment of the composition for oral care of thepresent invention, the calcium-containing component, thephosphorus-containing component, the calcium- and phosphorus-containingcomponent, the fluoro-containing component and the combination thereofthe present invention are adhered into the pores of the microparticles.

The materials constituting the microparticles of the present inventionare not limited, preferably may be composed of organic substance, or maybe composed of an oxide. For example, the preferred oxide constitutingthe microparticles includes silicon dioxide, silicoaluminate, titaniumdioxide, zinc oxide, aluminum oxide or a combination thereof. Morepreferably, the microparticles may be composed of silicon dioxide.

As to the microparticles composed of silicon dioxide, the source forproviding silicon dioxide is not limited, and may be an organic compoundor an inorganic compound containing silicon dioxide. More specifically,the silicon dioxide particles is preferably formed of silicate, waterglass or tetraalkoxy silane.

Moreover, the calcium-containing component of the present invention isnot limited, preferably may be selected from the group consisting ofcalcium oxide, calcium chloride, calcium carbonate, calcium nitrate,calcium hydroxide, calcium salt and a combination thereof.

The phosphorus-containing component of the present invention is notlimited, preferably may be selected from the group consisting ofphosphoric acid, sodium phosphate, phosphate, hydrogen phosphate,dihydrogen phosphate and a combination thereof.

The calcium- and phosphorus-containing component of the presentinvention is not limited, preferably may be selected from the groupconsisting of calcium phosphate, hydroxyapatite, tricalcium phosphate,tetracalcium phosphate, CaHPO₄, CaHPO₄.2H₂O, Ca₈H₂(PO₄)₆.5H₂O, Ca₂P₂O₇,Na₂O—CaO—SiO₂—P₂O₅ or a combination thereof.

The fluoro-containing component of the present invention is not limited,preferably may be selected from the group consisting of sodium fluoride,fluorides and salts thereof, and more preferably may be sodium fluoride.

Furthermore, the aforesaid components of the present invention are onlyused for exemplary illustration, and they are not intended to limit thescope of the present invention. That is, the composition of the presentinvention may further comprise an additional component other than theaforesaid components.

For example, similar to the aforesaid formulation for oral teeth of thepresent invention, the composition for oral care of the presentinvention may further comprise a fluoro-containing component. Thefluoro-containing component may be a fluoro-containing compound, such asa fluoro salt. Preferably, the fluoro-containing component is selectedfrom the group consisting of ammonium fluoride, calcium fluoride, sodiumfluoride, potassium fluoride, stannous fluoride, aluminum fluoride,sodium monofluorophosphate, sodium hexafluorosilicate and a combinationthereof. Further, the composition may further comprise a pH stabilizer.

In another preferred embodiment of the composition for oral care of thepresent invention, the pore diameters of the pores of the microparticlesare in a range of 2 nm to 50 nm, and the particle diameters of themicroparticles are in a range of 0.1 μm to 100 μm.

In another preferred embodiment of the composition for oral care of thepresent invention, the pH value of the composition for oral care of thepresent invention is not limited, and is preferably in a range of 2 to10, and more preferably in anyone of the ranges of 7 to 10, 4 to 7 or 5to 6 based on practical needs.

In addition, the method for preparing the composition for oral care ofthe present invention is not limited. For example, the method forpreparing the composition for oral care of the present invention may bemanufactured by sequentially using the sol-gel process to formmicroparticles having a plurality of pores, and using the impregnationprocess to allow a calcium-containing component, a fluoro-containingcomponent, a calcium- and phosphorus-containing component, afluoro-containing component or a combination thereof to adhered into thepores of the microparticle.

According to another aspect of the present invention, the presentinvention further provides a method for preventing or treating adentin-associated symptom or disease, which comprises a step ofadministering the aforesaid composition of the present invention to theoral cavity of a subject. More specifically, the dentin-associatedsymptom which can be prevented or treated by the aforesaid method ofadministering the composition of the present invention is not limited,preferably dentin hypersensitivity, crack tooth syndrome, enamel loss,dentin loss, cemento loss or postoperative hypersensitivity. On theother hand, the disease which can be prevented or treated by theaforesaid method of administering the composition of the presentinvention is not limited, preferably dental caries, root caries, toothfracture, root fracture, cervical abrasion, tooth wearing ordentin-associated pulp disease.

According to another aspect of the present invention, the presentinvention provides a product comprising the aforesaid composition of thepresent invention. The product includes a dental product, a toothpaste,a tooth powder, an ointment, a chewing gum, a chewable tablet, a troche,a mouthwash, a toothbrush or a strip. Preferably, the dental product mayinclude a dental cement, a dental bonding agent, a dental porcelainpowder, a pulp capping material, a retrograde filling material, a rootcanal filling material or a composite resin filling material.

Particularly, when a vital pulp therapy is performed, the aforesaid pulpcapping materials can be used to perform a direct or indirect cappingtreatment. Further, the aforesaid retrograde filling material may beused as a refilling material in apicoectomy .

EXAMPLE Example 1 Synthesis of Porous Microparticles Constituted bySilicon Dioxide

One gram of gelatin was added to 25 g of deionized water, placed in athermostatically controlled water batch at 40° C., and stirred for about15 minutes until complete dissolution was achieved, so as to form anaqueous gelatin solution.

Then, 4 g of sodium silicate was dissolved in 100 g of water, placed ina thermostatically controlled water batch at 40° C., and stirred forabout 2 minuets, to form an aqueous sodium silicate solution.

On the other hand, about 3.0 mL of 6M sulfuric acid was added to 100 gof water, and thoroughly stirred, to prepare an acidic solution haing apH value of about 1. The aqueous sodium silicate solution was mixed withthe acidic solution, to adjust pH to about 5. Then, sodium silicate isaged for 3 minutes, and the aqueous gelatin solution was added thereto.The mixture was placed in a thermostatically controlled water batch setat 40° C., and stirred for about 2 hours until complete dissolution wasachieved.

Subsequently, all of the mixture (containing the stock solution) waspoured into a polypropene (PP) hydrothermal bottle, and placed in anincubator set at 100° C. to undergo a hydrothermal reaction for one day.The obtained product was filtered, rinsed with water, and dried, toobtain a porous silicon dioxide material containing gelatin molecules.The organic matters were removed by calcining at 600° C. for 12 hours,to obtain silicon dioxide-based porous microparticles.

Furthermore, the size of the pores of the silicon dioxide material canbe controlled by adjusting the pH value of the solution and the numberof days of the hydrothermal reaction. Microparticles of different sizeswere prepared optionally by altering the source of silicon oxide and theweight ratios of silicon oxide and gelatin molecules.

Example 2 Preparation of Silicon Dioxide-Based Porous MicroparticlesContaining Calcium Carbonate

Zero point zero nine gram of oxalic acid was dissolved in 10 g ofde-ionized water, and 0.84 g of silicon carbonate was added thereto toform a first intermediate solution. Then, 10 g of alcohol was added tothe first intermediate solution, and stirred, and 0.5 g of the silicondioxide-based porous microparticles, as synthesized in example 1, wasadded thereto to form a second intermediate solution.

The aforesaid second intermediate solution was stirred until it driedup, and then placed in an incubator set at 100° C. for a day. Then, theresultant was taken out from the incubator, and placed into a furnaceventilated with air, with a temperature elevated from 200° C. to 400°C., and held at the temperature for 5 hours. Silicon dioxide-basedporous microparticles containing silicon carbonate (hereinafter,referred to as “Ca—Si based-porous particles”) were obtained. As shownin FIG. 1, the SEM observation of the Ca—Si based-porous particles ofthe example is provided.

It is noteworthy that the example is only exemplifying the contents ofthe present invention, not intended to limit the scope of the presentinvention. In other words, the method of the example can optionally beused in preparing porous microparticles comprising other component, inaddition to calcium carbonate. Further, the method for preparing silicondioxide-based porous microparticles containing calcium phosphate issubstantially the same as that of the present example, except that, inthe step of preparing the first intermediate solution, calcium carbonateis replaced with calcium phosphate.

Example 3 Preparation of a Formulation Comprising Ca—Si-Based PorousParticle

Eighty-five point seven percent of commercially available phosphoricacid (J.T. BAKER NALYZED, U.S.A.) was diluted to 31% of a phosphoricacid solution. Zero point zero five gram of the Ca—Si-based particlesobtained in example 2 was mixed with 0.15 mL of 31% of phosphoric acidsolution, to prepare a formulation comprising silicon-dioxide-basedporous microparticles containing calcium carbonate (hereinafter referredto as “CaCO3@pore SiO₂ formulation”).

Comparative Example 1 Seal & Protect® Formulation

A resin-type dentin bonding agent of commercially available Seal &Protect® (Densensitizer, Dentsply DeTrey, Konstanz, Germany) isprovided. Seal & Protect® is a light-curing and self-adhesive sealantcomposed of urethane dimethacrylate resin, polyrnerizabletrimethacrylate resins, dipentaerythritol pentaacrylate phosphate andamorphous silicon dioxide.

Test Example 1 Sealing Effect on Dentinal Tubules In Vitro

Twenty premolars and molars, provided with complete crowns having nocaries and no fillers, just removed from a human are collected.

An ultrasonic dental scaler (Sonicflex 2000, Kayo Co Biberbach, Germany)is used to remove dental calculus and periodontal tissues from thepremolars and molars. Then, the premolars and molars are stocked in 4°C. distilled water, so as to maintain the freshness of the dentin.

Before applying the formulation, the teeth are taken out of the water,and the enamel at the occlusion site is removed in a horizontaldirection using a low speed saw (Isomet low speed saw, Buehler, LTD.),and incised at a distance of 1.5 mm along the direction of the neck toobtain a specimen of dentin. Then, a tapered fissure but (1961 taperedfissure but) is used to create a groove on the back of the experimentalarea of each of the specimens, to guide the direction of future incisionof the specimens. Thirty-seven point five percents of phosphoric acid asgel etchant (Kerr Co USA) is used to acid etch the specimens up to 40seconds. Then, a large amount of distilled water is used to wash thecoating layer, and the surfaces of the specimens are blow-dried.

The formulation of Example 3 and Seal & Protect® of Comparative example1 are used, respectively, by the following approaches, to coat thespecimens.

As to CaCO₃@porous SiO₂ formulation of Example 3, the formulation iscoated and pressed tightly on the surfaces of the specimens using asmall brush. After 3 minutes, the formulation on the surfaces is rinsedwith water. Then, the above step is repeated for three times, whereinalkaline water with pH of 9 is used at the third time to wash theformulation on the surfaces,

As to Seal & Protect® of comparative example 1, Seal & Protect® iscoated on the surfaces of tooth samples, and allowed it to react for 20seconds. Then, the tooth samples are blow-dried for 5 seconds, andilluminated for 10 seconds. Subsequently, Seal & Protect® is coated onthe surfaces of the tooth samples, blow-dried for 5 seconds, andilluminated for 10 seconds. The coating of Seal & Protect® is completed.

Finally, a field emission scanning electronic microscope (SEM; FieldEmission Scanning Electronic Microscope Hitachi S-800, Hitachi Co.,Tokyo, Japan) is used to observe the depth of the precipitates in thedentinal tubules in each of the specimens.

FIGS. 2a and 2b show the SEM observations of each of the specimens,respectively. Mores specifically, FIG. 2a shows the SEM observation ofdentin specimens coated with CaCO₃@porous SiO₂ formulation, and FIG. 2bshows the SEM observation of dentin specimens coated with Seal &Protect® formulation.

After comparison, it is found that, in the dentinal tubules, the depthsof the precipitates provided by Seal & Protect® of Comparative exampleare only about 10 μm. There are still existed gaps between theprecipitates and the walls of the dentinal tubules, and cannotcompletely seal the dentinal tubules (please refer to FIG. 2b ).However, the depths of the precipitates provided by the formulation ofExample 3 are as deep as about 60 μm, and the dentinal tubules arealmost 100% sealed. In other words, the CaCO₃@porous SiO₂ formulation ofExample 3 can provide better sealing effect to dentin.

Test Example 2 Permeability Test on Dentinal Tubules In Vitro

The permeability exhibited on the dentin coated with the formulation ofexample 3 and Seal & Protect® of comparative example, respectively, isevaluated using a flow model test. The lower the determined permeabilityis, the better the sealing effect of dentin is.

As to the operation of the liquid module test, a glass tube is firstlyprovided. One end of the glass tube is sealed with a dentin specimen,while the other end of the glass tube is provided with 0.15 g/cm² of apressure source. A air bubble is formed in the glass tube. After 72hours, the distance that the air bubble moves is measured, and theresult is used as a baseline. Then, the dentin specimens are coated,while the pressure is continuously provided. After 72 hours, thedistance moved that the air bubble moves is measured again, and theresult obtained is the permeability distance. The base line andpermeability distance are substituted into the following equation, tocalculate the permeability of dentin.

Permeability (%)=baseline/permeability distance×100%

As to the step of coating the dentin specimen, the step is variedaccording to the coating material.

More specifically, the formulation of Example 3 is coated to thesurfaces of the dentin specimens, and then pressed tightly. After 10minutes, the surfaces of the specimens are rinsed with water.

Seal & Protect® of Comparative example 1 is coated on the surfaces oftooth samples, and allowed them to react for 20 seconds. Then, the toothsamples are blow-dried for 5 seconds, and illuminated for 10 seconds.Next, Seal & Protect® is coated again on the surfaces of the toothsamples. Then, the tooth samples are blow-dried for 5 seconds, andilluminated for 10 seconds. The results showed that the permeability thedentin specimens of Comparative example 1 exhibit are about 40%, whereasthe dentin specimens of example 3 are about 15%. That is to say, theformulation of Example 3 of the present invention provided bettersealing effect of dentin.

Test Example 3 Sealing Effect on Dentinal Tubules in an Animal Test

Dogs are anesthetized. A cavity with a length of about 5 mm, a width ofabout 3 mm, and a depth of about 1.5 mm to 2 mm is created on the upperand lower canine on both sides and the neck portion near the first molarof the dog, using a high-speed hand piece equipped with a spray nozzlefor dental use. Then, 37.5% of phosphoric acid gel etchant for dentaluse (Kerr Co. CA, USA) is used for acid etching for 40 seconds, toremove the coating layers. Then, a large amount of water is used to washthe gel etchant away from the surfaces of the teeth, so as to form toothsamples to be filled.

The formulations of Example 3 and Comparative example 1 are used,respectively, to fill the tooth samples by the following approaches.

As to the formulation of Example 3, the formulation is coated andpressed tightly on the surfaces of the tooth samples with dentin exposedfrom the cavity. After 3 minutes, the coated tooth samples are swabbedusing a cotton ball dipped in distilled water. Then, the aforesaidswabbing step is repeated for three times, wherein swabbing is performedusing a cotton ball dipped with alkaline water at pH 9 in the thirdtime. Then, a self-polymeric glass ionomer (GC Fuji II™, GC, Tokyo,Japan) is used to fill the cavity.

Seal & Protect® of Comparative example 1 is coated on the surfaces ofthe tooth samples, and allowed to react for 20 seconds. Then, thesamples are blow-dried for 5 seconds, and illuminated for 10 seconds.Seal & Protect® is coated again on the surfaces of the tooth samples.Then, the tooth samples are blow-dried for 5 seconds, and illuminatedfor 10 seconds. After the completion of the coating of Seal & Protect®,the self-polymeric glass ionomer (GC FujiII™, GC, Tokyo, Japan) is usedto fill the cavity.

After 1 week, the tooth samples from each of the groups are removed.Then, the tooth samples are incised to prepare specimens. The depths ofthe precipitates in the dentinal tubules in the specimens of each of thegroups are observed using a field emission scanning electronicmicroscope (Hitachi S-800, Hitachi Co., Tokyo, Japan).

FIGS. 3a and 3b show the observations of specimens of each of thegroups. FIG. 3a shows the SEM observation of the dentin samples of alive dog coated with the CaCO₃@porous SiO₂ formulation of Example 3, andFIG. 3b shows the SEM observation of the dentin samples of a live dogcoated with Seal & Protect® of Comparative example 1.

After comparison, it is found that, in the dentinal tubules, Seal &Protect® of Comparative example 1 coated a resin layer on the dentinsurfaces, and the resin layer does not attach tightly with the dentinsurfaces. The depth of the resin extending into the dentinal tubules isonly about 5 μm (please refer to FIG. 3b ). The formulation of Example 3provides precipitates with depths of about 40 μm, and the dentinaltubules are almost completely sealed (please refer to FIG. 3a ). Inother words, the CaCO₃@porous SiO₂ formulation of example 3 can providebetter sealing effect to dentin.

Test Example 4 Evaluation of Pulp Irritation

Pulp irritation is evaluated in conformity to the ISO-7405 pulpirritation test.

Firstly, dogs are anesthetized. A cavity with a length of about 5 mm, awidth of about 3 mm, and a depth of about 1.5 mm to 2 mm is created onthe upper and lower canine on both sides and the neck portion near thefirst molar of the dog, using a high-speed hand piece equipped with aspray nozzle for dental use. Then, 37.5% of phosphoric acid gel etchantfor dental use (Kerr Co. CA, USA) is used for acid etching for 40seconds, to remove the coating layers. Then, a large amount of water isused to wash the gel etchant away from the surfaces of the teeth, so asto form tooth samples to be filled.

The formulations of Example 3 and comparative example 1 are used,respectively, to fill the tooth samples by the following approaches.

As to the formulation of Example 3, the formulation is coated andpressed tightly on the surfaces of the tooth samples with dentin exposedfrom the cavity. After 3 minutes, the coated tooth samples are swabbedusing a cotton ball dipped in distilled water. Then, the aforesaidswabbing step is repeated for three times, wherein swabbing is performedusing a cotton ball dipped with alkaline water at pH=9 in the thirdtime. Then, a polymeric glass ionomer (GC Fuji II™, GC, Tokyo, Japan) isused to fill the cavity.

Seal & Protect® of Comparative example 1 is coated on the surfaces ofthe tooth samples, and allowed them to react for 20 seconds. Then, thesamples are blow-dried for 5 seconds, and illuminated for 10 seconds.Seal & Protect® is coated again on the surfaces of the tooth samples.Then, the tooth samples are blow-dried for 5 seconds, and illuminatedfor 10 seconds. After the completion of the coating of Seal & Protect®,a self-polymeric glass ionomer (GC FujiII™, GC, Tokyo, Japan) is used tofill the cavity.

On days 7, 28 and 70, the tooth samples of each of the groups to befilled is incised to be made into test slices for pulp irritation, inconformity to ISO-7405 pulp irritation test. Then, an optical microscopeis used to observe the levels of inflammation of the cells in the pulpcavity of the slices of each of the groups.

FIG. 4a and FIG. 4b show the observations of the levels of inflammationof the cells in the pulp cavity of the slices of each of the groups onday 7. More specifically, FIG. 4a shows the observation of the pulptissue morphology of the dentin specimens of a live dog coated with theCaCO₂@porous SiO₂ formulation of Example 3, and FIG. 4b shows theobservation of the pulp tissue morphology of the dentin specimens of alive dog coated with Seal & Protect® of comparative Example 1.

As shown in FIG. 4b , moderate inflammation is observed in the testslices filled with Seal & Protect® of Comparative example 1. Morespecifically, it is observed that the cell densities in the pulp tissuesare elevated, lymphocytes are infiltrated, and the blood vessels arefilled with red blood cells.

By contrast, as shown in FIG. 4a , the test slices filled with theCaCO₃@porous SiO₂ formulation of Example 3 showed no inflammation ormild inflammation. More specifically, the pulp tissues had the samemorphology as that of the normal pulp tissues. There are no inflamedcells, or scarce inflamed cells. No angiogenesis or diapiresis areobserved in the pulp tissues.

It is known from the results of test example 3 that, the CaCO₃@porousSiO₂ formulation of Example 3 provided stronger anti-inflammationability for a long period. Thus, the CaCO₃@porous SiO₂ formulation ofexample 3 can provide a therapeutic effect at long-term.

The invention has been described using exemplary preferred embodiments.However, it is to be understood that the scope of the invention is notlimited to the disclosed embodiments. On the contrary, it is intended ocover various modifications and similar rearrangement. The scope of theclaims therefore should be accorded the broadest interpretation so as toencompass all such modifications and similar arrangements.

1-65. (canceled)
 66. A method for preventing or treating adentin-associated symptom or disease in a subject in need thereof,comprising administering a formulation to an oral cavity of the subject,wherein the formulation comprises: a plurality of calcium ion carrierswhich are microparticles having a plurality of pores; a plurality ofcalcium-containing particulates composed of calcium carbonate, calciumchloride, calcium nitrate or a combination thereof, wherein thecalcium-containing particulates are carried by the calcium ion carriersand adhered into the plurality of pores of the microparticles; and anacidic solution for releasing calcium ions from the calcium-containingparticulates carried by the calcium ion carriers for formingprecipitation deep in dentinal tubules.
 67. The method of claim 66,wherein the dentin-associated symptom comprises dentin hypersensitivity,crack tooth syndrome, enamel loss, dentin loss, cemento loss, orpostoperative hypersensitivity.
 68. The method of claim 66, wherein thedentin-associated disease comprises dental caries, root caries, toothfracture, root fracture, cervical abrasion, tooth wearing, ordentin-associated pulp disease.
 69. The method of claim 66, wherein thecalcium ion carriers are composed of at least an inorganic compoundselected from the group consisting of silicon dioxide, silicoaluminate,titanium dioxide, zinc oxide, aluminum oxide and a combination thereof.70. The method of claim 66, wherein the microparticles have particlediameters in a range of from 0.1 μm to 100 μm.
 71. The method of claim66, wherein the plurality of pores have pore diameters in a range offrom 1 nm to 100 nm.
 72. The method of claim 71, wherein the porediameters of the plurality of pores are in a range of from 2 nm to 50nm.
 73. The method of claim 66, wherein the acidic solution is at leastone selected from the group consisting of a phosphoric acid solution, anoxalic acid solution and a citric acid solution.
 74. The method of claim73, wherein the acidic solution is the phosphoric acid solution.
 75. Themethod of claim 74, wherein the phosphoric acid solution has aconcentration in a range of from 1% to 65%.
 76. The method of claim 75,wherein the concentration of the phosphoric acid solution is in a rangeof from 25% to 45%.
 77. The method of claim 76, wherein theconcentration of the phosphoric acid solution is in a range of from 31%to 40%.
 78. The method of claim 66, wherein a ratio of the volume of theacidic solution to the weight of the calcium ion carriers carrying thecalcium-containing particulates is in a range of from 1 mL/g to 10 mL/g.79. The method of claim 78, wherein the ratio of the volume of theacidic solution to the weight of the calcium ion carriers carrying thecalcium-containing particulates is in a range of from 2 mL/g to 5 mL/g.80. The method of claim 66, wherein the formulation is present as a gel,a paste, a slurry, an emulsion or a glue.
 81. The method of claim 66,wherein the formulation has a pH value in a range of from 4 to
 6. 82.The method of claim 66, wherein the formulation has a pH value in arange of from 5 to
 6. 83. The method of claim 66, wherein theformulation further comprises at least one of a phosphorus-containingcomponent and a fluorine-containing component.
 84. The method of claim83, wherein the phosphorus-containing component is at least one selectedfrom the group consisting of phosphate, hydrogen phosphate, dihydrogenphosphate, hydroxyapatite, sodium phosphate, calcium phosphate,tricalcium phosphate, tetracalcium phosphate, CaHPO₄, Ca₈H₂(PO₄)₆.5H₂O,Ca₂P₂O₇, Na₂O—CaO—SiO₂—P₂O₅ and any combination thereof.
 85. The methodof claim 83, wherein the fluorine-containing component is at least oneselected from the group consisting of ammonium fluoride, calciumfluoride, sodium fluoride, potassium fluoride, stannous fluoride,aluminum fluoride, sodium monofluorophosphate, sodium hexafluorosilicateand any combination thereof.